Beads bound to a solid support and to nucleic acids

• US RE44,693 E1
• Filed: 07/22/2009
• Issued: 01/07/2014
• Est. Priority Date: 11/06/1996
• Status: Expired due to Term

• Current Assignee: Agena Bioscience Inc. (Mesa Laboratories Inc.)
• Original Assignee: Sequenom Incorporated (Laboratory Corporation Of America Holdings)
• Inventors: Lough, David M., Kster, Hubert
• Primary Examiner(s): MAIER, LEIGH C
• Application Number:
  US12/507,124
• Time in Patent Office: 1,630 Days
• US Class Current: 435/6.11
• CPC Class Codes: B01J 2219/00315 Microtiter plates
• CPC Class Codes: B01J 2219/00317 Microwell devices, i.e. having large numbers of wells
• CPC Class Codes: B01J 2219/00387 Applications using probes
• CPC Class Codes: B01J 2219/00468 by manipulation of individual beads
• CPC Class Codes: B01J 2219/00497 Features relating to the solid phase supports
• CPC Class Codes: B01J 2219/005 Beads
• CPC Class Codes: B01J 2219/00504 Pins
• CPC Class Codes: B01J 2219/00511 Walls of reactor vessels
• CPC Class Codes: B01J 2219/0052 in the shape of elongated tubes
• CPC Class Codes: B01J 2219/00527 Sheets
• CPC Class Codes: B01J 2219/00585 Parallel processes
• CPC Class Codes: B01J 2219/00596 Solid-phase processes
• CPC Class Codes: B01J 2219/00648 by the use of solid beads
• CPC Class Codes: B01J 2219/00659 Two-dimensional arrays
• CPC Class Codes: B01J 2219/0072 Organic compounds
• CPC Class Codes: B01J 2219/00722 Nucleotides
• CPC Class Codes: B01J 2219/00725 Peptides
• CPC Class Codes: C07H 21/00 Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids
• CPC Class Codes: C12Q 1/6837 using probe arrays or probe chips C12Q1/6874 takes precedence
• CPC Class Codes: C12Q 1/6872 involving mass spectrometry
• CPC Class Codes: C40B 40/06 : Libraries containing nucleotides or polynucleotides, or derivatives thereof
• CPC Class Codes: C40B 40/10 : Libraries containing peptides or polypeptides, or derivatives thereof
• CPC Class Codes: C40B 60/14 : for creating libraries
• CPC Class Codes: G01N 2035/1069 : by adjusting the spacing between multiple probes of a single transferring head
• CPC Class Codes: G01N 35/1067 : for transfer to or from containers having different spacing

Abstract

Novel compositions comprised of at least one bead conjugated to a solid support and further conjugated to at least one nucleic acid and preferred methods for making the novel compositions are described. As compared to “flat” surfaces, beads linked to a solid support provide an increased surface area for immobilization of nucleic acids. Furthermore, by selecting a bead with the desired functionality, a practitioner can select a functionalization chemistry for immobilizing nucleic acids, which is different from the chemistry of the solid support.

Claims

1. A composition, comprising a bead conjugated to a solid copper support by covalent attachment and further conjugated to a nucleic acid, wherein the solid support is selected from the group consisting of multiwell plates, arrays of pits and multiwell supports comprising nanoliter wells wherein the bead is bound to the solid support and the nucleic acid is bound to the bead using orthogonally cleavable linkers.

2. A composition of claim 1, wherein the bead is made from a material selected from the group consisting of:

silica gel, glass, magnet, 4-(hydroxymethyl)phenoxymethylcopoly(styrene—

1% divinylbenzene) resin, chloromethylated copolystyrene—

divinylbenzene resin, metal, plastic, cellulose, dextran cross-linked with epichlorohydrin, and agarose.

3. A composition of claim 1, wherein the bead is swellable.

4. A composition of claim 1, wherein the bead is nonswellable.

5. A composition of claim 1, wherein the bead is in the range of 1 to 100 μ

m in diameter.

6. A composition of claim 1, wherein the nucleic acid is DNA.

7. A composition of claim 1, wherein the nucleic acid is RNA.

24. The composition of claim 1, wherein the conjugation of the bead to the nucleic acid comprises a spacer.

25. The composition of claim 1, wherein the conjugation of the bead to the nucleic acid comprises streptavidin-biotin DNA.

26. The composition of claim 1, wherein the bead can be selectively cleaved from the solid surface without cleaving the nucleic acid from the bead.

27. The composition of claim 1, wherein the conjugation of the bead to the solid surface comprises a disulfide linker.

8. A process of making a bead conjugated to a solid support and further conjugated to a nucleic acid, comprising the steps of conjugating a bead to a nucleic acid;

and conjugating a bead to a solid support, wherein the solid support is selected from the group consisting of multiwell plates, arrays of pits, and multiwell supports comprising nanoliter wells.

9. A process of claim 8, wherein the bead is functionalized.

10. A process of claim 9, wherein the bead is functionalized with carboxy functional groups.

11. A process of claim 9, wherein the bead is functionalized with amino functional groups.

12. A process of claim 9, wherein the bead is conjugated to the nucleic acid prior to conjugation of the bead to the solid support.

13. A process of claim 9, wherein the bead is conjugated to the nucleic acid after the bead is conjugated to the solid support.

18. The method of claim 8, wherein conjugation is effected with a crosslinking agent.

14. A kit, comprising:

i) beads,

ii) an insoluble support, and

iii) conjugation means for linking nucleic acids to the beads and the beads to the support.

15. The kit of claim 14, wherein the solid support is selected from the group consisting of:

beads, capillaries, plates, membranes, wafers, combs, pins, wafers with arrays of pits, and supports with nanoliter wells.

16. The kit of claim 14, wherein the bead is made from material selected from the group consisting of silica gel, glass, magnet, p-benzyloxybenzyl alcohol copolystyrene-divinyl benzene (DVB) resin, chlorotritylchloride copolystyrene-DVB resin, chloromethylated copolystyrene-DVB resin, metal, plastic, cellulose, cross-linked dextran, and agarose gel.

17. A composition, comprising a bead conjugated to a solid support and further conjugated to a nucleic acid, wherein conjugation is effected with a crosslinking agent.

19. A composition, comprising a bead conjugated to a solid support and further conjugated to a nucleic acid molecule comprising protein nucleic acid.

20. A composition, comprising a bead conjugated to a solid support and further conjugated to a nucleic acid, wherein conjugation is effected through a photocleavable linkage.

21. The composition of claim 20, wherein the linkage is cleaved by exposure to a laser.

22. The composition of claim 20, wherein the linkage is cleaved by exposure to electromagnetic radiation selected from ultravioltet, visible, infrared radiation or electromagnetic radiation generated by fluorescence or chemiluminescence, or combinations thereof.

23. A composition, comprising a bead conjugated to a solid support and further conjugated to a nucleic acid, wherein conjugation is effected through ionic linkages.